LOCUS       sequence-363957-        6571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS2_PARRC_98-130 attached to 2xFKBP. The FKBP can be used for 
            inducible dimerization and condensate formation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6571)
  AUTHORS   Veling MT, Nguyen DT, Thadani NN, Oster ME, Rollins NJ, Brock KP, 
            Bethel NP, Lim S, Baker D, Way JC, Marks DS, Chang RL, Silver PA
  TITLE     Natural and Designed Proteins Inspired by Extremotolerant Organisms 
            Can Form Condensates and Attenuate Apoptosis in Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1292-1302. doi: 
            10.1021/acssynbio.1c00572. Epub 2022 Feb 18.
  PUBMED    35176859
REFERENCE   2  (bases 1 to 6571)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.1c00572"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1292-1302"
COMMENT     Sequence Label: CAHS2_PARRC_98-130_mCh-2xFKBP (pBS1120)
FEATURES             Location/Qualifiers
     source          1..6571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    826..844
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    847..865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     875..899
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1008..1017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1857..2177
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2184..2504
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2511..2537
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2585..2602)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2591..2815
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2861..3289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2948..2967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3158..3179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3298..3318)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3303..3632
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3483..3618
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3545..3564
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3680..3727
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3746..4120
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4250..4371
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4287..4306)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4341..4360
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4420..4436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4444..4460
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4468..4474,4475..4492,4493..4498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4513..4534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4651..4668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4822..5410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4902..4921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5581..6372,6373..6441))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMAVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6204..6223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6442..6546)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      841 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      901 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      961 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccgcc accatggccg
     1021 ccaagactag agagaacgag atgctgggcc agcagtacga gaaagagttg gagcggaagt
     1081 ccgaggctta cagaaaacac caggaggtgg aggcctccgg aggcggcgga tccggcggaa
     1141 tggtgtctaa aggcgaggag gataacatgg ctataatcaa ggaatttatg aggttcaagg
     1201 tgcatatgga gggatctgtg aacggtcacg agttcgaaat cgagggcgaa ggcgaggggc
     1261 gaccctatga agggacacag acggccaaac tgaaggtgac caagggtggc cccctgcctt
     1321 tcgcctggga catcctgtca ccccagttta tgtacggctc taaggcttac gttaagcacc
     1381 ctgccgatat tccggactac ttgaaactga gcttccctga agggttcaag tgggaacggg
     1441 taatgaattt tgaggatggc ggtgtagtca cagttaccca ggacagctct cttcaggacg
     1501 gagaatttat ctataaggtt aaacttcggg gcactaactt cccatcagac ggccccgtga
     1561 tgcagaagaa aactatgggc tgggaagcca gctcagaacg catgtatccc gaggacggag
     1621 cactgaaggg agaaatcaag cagcgactca agctgaagga tgggggacat tatgatgctg
     1681 aggtcaaaac cacctataag gccaagaaac ccgttcagct tcctggggca tacaacgtga
     1741 atatcaaact ggatattaca tcccataacg aagactatac catcgtggaa cagtacgagc
     1801 gggccgaggg cagacatagc acagggggca tggatgaatt gtacaaggcg actagaggcg
     1861 tccaagtcga aaccattagt cccggcgatg gcagaacatt tcctaaaagg ggacaaacat
     1921 gtgtcgtcca ttatacaggc atgttggagg acggcaaaaa ggtggacagt agtagagatc
     1981 gcaataaacc tttcaaattc atgttgggaa aacaagaagt cattagggga tgggaggagg
     2041 gcgtggctca aatgtccgtc ggccaacgcg ctaagctcac catcagcccc gactacgcat
     2101 acggcgctac cggacatccc ggaattattc cccctcacgc taccttggtg tttgacgtcg
     2161 aactgttgaa gctcgagact agaggagtgc aggtggagac tatctcccca ggagacgggc
     2221 gcaccttccc caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg
     2281 gaaagaaagt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     2341 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     2401 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     2461 cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaactagt tatccgtacg
     2521 acgtaccaga ctacgcataa gcggccgctc gagtctagag ggcccgttta aacccgctga
     2581 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2641 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2701 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2761 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2821 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3061 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3181 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3301 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3361 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3421 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3541 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3601 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3661 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     3721 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     3781 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3841 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3901 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     3961 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     4021 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     4081 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     4141 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4201 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4261 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4321 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4381 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4441 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4501 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4561 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4681 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4981 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5221 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5521 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5581 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5641 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5701 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5761 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5821 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5881 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5941 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6001 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6061 catggttatg gcagcactgc ataattctct tactgccatg ccatccgtaa gatgcttttc
     6121 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6181 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6241 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6301 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6361 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6421 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6541 tccgcgcaca tttccccgaa aagtgccacc t
//
